Escherichia coli BL21|356365 |BNCC

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Escherichia coli BL21

Subculture procedure	① Prepare 1-2 of the above plates; ② Open the safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	37 ℃; 18-24h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G - (red), Bacillus, purity: pure
Sharing mode	Public welfare sharing

BL21 Chemically Competent Cell

Storage conditions: -20 ℃

Validity period: 5 years

No. 356365

Product format: bacteria tablets, vials

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Usage: competent cells, plasmids, genetic engineering

Notes: this product is transported at room temperature. please store it at -20 ℃ after receiving it. If any abnormal situation such as damage is found on the same day, please contact customer service within 24 hours. The Xilin bottle shall be used up once after opening and shall not be retained. Please operate in strict accordance with this instruction, otherwise the replacement service will not be provided if the strain is abnormal and inactivated.

Medium:LB agar medium, yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, agar 15.0g, distilled water 1.0 L,pH 7.0. Sterilization at 121 ℃ for 15min.

Growth conditions

Temperature:37 ℃

Atmosphere: Aerobic

Instruction

A. Recovery

1. Prepare 2 LB plates;

2. Under aseptic environment, 0.5mL of liquid culture medium is injected into the penilin bottle to fully dissolve the bacteria tablets;

3. Scribe the suspension on the plate, 2 plates, 18-24h to grow a single colony;

B. Competent preparation (for reference)

1. Escherichia coli DH5α Pick a single colony on the plate and connect it to a test tube of 3ml LB liquid medium, and culture at 37 ℃ for overnight.

2. Take 1ml of bacterial solution and transfer it to a conical flask containing 50ml LB liquid culture medium, shake and culture at 37 ℃ for 2~3h,A600 should be between 0.4~0.5, and the number of cells must be <10⁸CFU/ml.

3. Transfer 1ml of bacterial liquid to a 1.5 ml centrifuge tube with a pipette gun and place it on ice for 10min.

4. Centrifugation for 10min,4000r /min.

5. Pour out the culture solution and use cold 0.1mol/l CaCl₂ 200µl suspension precipitation, immediately placed on ice for 30min.

6. Low temperature centrifugation for 10min,4000r /min, cells were recovered.

7. Use ice-cold 0.1mol/l CaCl₂ 200µl suspension cells.

8. Subpackage cells, each 200ul, this cell is competent cells.

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